Meteorological factors affecting daily urticaceae pollen counts in southwest Spain

The influence of meteorological factors on daily Urticaceae pollen counts were studied in Córdoba (southwest Spain) in 1996 and 1997. The daily Urticaceae pollen concentrations were obtained by using a Hirst-type volumetric sampler, and meteorological data were obtained from the Córdoba airport, located near the sampling site. The highest correlation between pollen concentration and meteorological parameters was obtained during non-rainy seasons. Temperature was found to be the most important meteorological parameter influencing pollen counts in spring, as temperature is the main reason for the increase of pollen concentration in the atmosphere. In autumn, humidity was another important parameter influencing pollen counts. Rain, however, did not appear to be significant. The influence of the pollen concentration of the 2 previous days and the pollen concentration of the previous day has been studied. During periods with low precipitation, the pollen concentration of the previous day was a useful predictor of Urticaceae pollen concentrations for the following day. Received: 4 January 1999 / Revised: 26 July 1999 / Accepted: 6 September 1999     

Analysis of spatial and yearly variation in winter survival of winter wheat

 Four years of winter survival data for winter wheat (Triticum aestivum L.) were collected on a loam soil located on the Central Experimental Farm at Ottawa, Ontario (45° 23′N, 75° 43′W). The site was low-lying and subject to frequent winter flooding and ice sheet formation. It appeared level although there was microtopographic variation with a range in elevation of approximately 0.15 m. The objective of the study was to gain insight into factors which might affect winter survival. Two varieties, a soft white and a hard red winter wheat, were planted in September. Crop establishment was measured in late fall and the percentage survival was measured in April of the following year. We assumed the large spatial differences in survival were not totally random, but rather were affected by spatial variation in environmental factors such as snow and ice depth, soil moisture and temperature. Hourly measurements of soil temperature at a depth of 0.05 m were recorded throughout the fall, winter and spring. Fall and spring soil moisture at the same depth were measured on the plot, as well as snow and ice depth at selected times throughout the winter. Measurements were taken on a grid covering the plot to help explain spatial variation in survival. In addition, detailed measurements of elevation were taken on a grid. Meteorological data were available from the nearby weather site. While soil temperatures were never low enough to kill plants according to the CERES model, the spatial variation in winter survival was associated with differences in elevation and the resulting surface drainage patterns. Received: 23 March 1998/Accepted: 21 October 1998     

Spectrophotometric analysis as a complementary technique to aerobiology in the study of solid particles in the air

Hitherto, aerobiology has mainly focused on studying and analysing the biologic material found in the atmosphere (pollen grains and fungal spores), due to its allergologic and agricultural interest. However, the increase in respiratory tract problems caused by other solid particles suspended in the air has made studies including biologic and non-biologic material more frequent. This paper describes a simple technique based on spectrophotometric analysis. The results thus obtained can complement the outcome obtained by traditional aerobiologic techniques if the method is used properly and the equipment properly calibrated and an idea of the solid particles in the air can be obtained. The combination of both aerobiological and spectrophotometric analysis makes it possible to estimate the concentration of solid-particle material in the air, and we can also estimate the percentage of pollen grains and fungal spores among the general spectrum of particles. The results also show that a great part of the solid particulates originate from human activities. However, it was also observed that they emerge from natural sources as such as fungi and higher plants.     

Bioclimatic factors affecting daily Cupressaceae flowering in southwest Spain

 Daily variations in Cupressaceae pollen counts were analysed, together with climatic variables in the southwest of Spain, in order to understand the bioclimatic variables affecting Cupressaceae flowering in a Mediterranean climate. The study was carried out using the Spanish Aerobiology Network recommendations, using a Burkard Hirst-type spore trap. Regression analyses were applied to the pollen counts of the winter months of 13 consecutive years. The regression analysis applied to each year indicated that temperature is the most important variable to include in the model. Temperature affected the pollen counts differently according to the yearly climatic trend. During the coldest years, pollen counts depended on the minimum temperature, while in the warmest ones they were affected by higher temperatures. However, during the driest years, pollen counts depended more on the humidity level, while in the wettest years they depended more on the hours of sunshine. Received: 12 November 1996 / Accepted: 19 August 1997     

Source of Bet v 1 loaded inhalable particles from birch revealed

 Allergenic proteins present in pollen grains, when inhaled, interact with the airways to cause an attack of asthma in susceptible humans. In one system, grass pollen grains rupture osmotically in rainfall, releasing allergen-containing inhalable particles into the atmosphere. In contrast, birch tree pollen grains do not rupture under these conditions, yet the major allergen, Bet v 1, has been detected in the atmosphere in inhalable particles of unknown origin. It is possible that Bet v 1 may diffuse from intact settled pollen grains and the allergenic material may again become airborne, interacting with settled fine particles from other sources prior to resuspension. This study investigates the mechanism for the release of birch pollen allergen-containing inhalable particles from pollen grains. We propose the hypothesis that (1) airborne birch pollen grains settle on nearby leaf surfaces; (2) then, following light rainfall, the grains germinate and, (3) later, pollen tubes burst, releasing inhalable particles carrying Bet v 1 into the atmospheric aerosol.   We used microscopic analyses of pollen behaviour following anther opening, a Burkard volumetric trap for pollen counts and a high volume air sampler with a two-stage cascade impactor for quantitative immunochemical analyses of Bet v 1. On dry days of high birch pollen count (48 grains/m³, 1.5 ng/m³ of Bet v 1), we found that the surfaces of birch leaves became coated with pollen. This ”pollen rain” is a source of secondary emission of allergens into the atmosphere. We observed that following light rainfall (<1 mm per day), about 80% of the birch pollen grains germinated, producing pollen tubes, especially in the sticky surface secretions of leaf glands. These pollen tubes may grow up to 300 μm in length prior to rupturing, each releasing about 400 starch granules coated with allergen molecules that may, after drying, be dispersed into the aerosol. On these days following light rainfall, the highest atmospheric levels of Bet v 1 (1.18 ng/m³) are associated with inhalable particles. Following heavy rainfall, both pollen and inhalable particles are washed from the atmosphere. Immunoprinting studies show that Bet v 1 is associated with starch granules rather than the smaller orbicules. Bet v 1 is present in the atmosphere in large particles, i.e. in particular pollen grains and in inhalable particles, i.e. in particular starch granules. Received: 28 May 1997 / Revision accepted: 18 August 1997     

Unreduced diploid nuclei in Cupressus dupreziana A. Camus pollen

Size and DNA content of pollen of Cupressus dupreziana A. Camus, a highly endangered Mediterranean conifer, were assessed by cytomorphological observations and flow cytometric analyses and then compared to C. sempervirens L. pollen. Mature C. dupreziana pollen was composed of two uninucleated types of pollen grains differing in size. Around 35% of the grains exhibited a size similar to C. sempervirens pollen, while 65% exhibited a larger diameter. However only one peak of fluorescence was detected by flow cytometry. DNA content of C. dupreziana pollen was twice the DNA content of C. sempervirens pollen. Comparison of DNA contents of mature and germinating pollen revealed that mature pollen of both species were arrested in the G2 phase. Comparison with the DNA content of somatic tissue (2C) provided evidence for the production of unreduced pollen in C. dupreziana. This unexpected feature in gymnosperms is discussed in terms of reproductive strategy of this species. Received: 11 December 1999 / Accepted: 22 December 1999     

Analytical approximations for the orientation distribution of small dipolar particles in steady shear flows

 Analytic approximations are obtained to solutions of the steady Fokker-Planck equation describing the probability density functions for the orientation of dipolar particles in a steady, low-Reynolds-number shear flow and a uniform external field. Exact computer algebra is used to solve the equation in terms of a truncated spherical harmonic expansion. It is demonstrated that very low orders of approximation are required for spheres but that spheroids introduce resolution problems in certain flow regimes. Moments of the orientation probability density function are derived and applications to swimming cells in bioconvection are discussed. A separate asymptotic expansion is performed for the case in which spherical particles are in a flow with high vorticity, and the results are compared with the truncated spherical harmonic expansion. Agreement between the two methods is excellent. Received 10 February 1997; received in revised form 26 May 1997     

Transglutaminase-catalyzed modification of cytoskeletal proteins by polyamines during the germination of Malus domestica pollen

 We investigated polyamine linkage to different structural proteins in pollen of Malus domestica Borkh. cv Red Chief at different phases of germination. This linkage has the characteristics of covalent linkages, indeed, it could be catalyzed by transglutaminase (TGase; EC 2.3.2.13). This assumption is supported by: (1) formation of a labelled TCA pellet and selective labelling of endogenous proteins by covalent binding of radioactive polyamines and (2) cross-reactivity of two different polyclonal antibodies against mammalian TGases; western blot analysis allowed us to detect a protein of about 80 kDa in both rehydrated ungerminated and germinated pollen. TGase activity was high at 90 min after germination and was influenced by Ca²⁺ supply only in the rehydrated ungerminated pollen. Extraction by Triton X-100 suggests that pollen TGase was at least partially membrane-bound. The enzyme catalyzed the incorporation of polyamines mainly into proteins having a molecular mass of 43 kDa and 52–58 kDa in both ungerminated and germinated pollen. These bands matched immunolabelled spots identified by mouse monoclonal anti-actin and anti-α-tubulin antibodies. Supplying exogenous actin and tubulin in a cell-free extract of rehydrated ungerminated and germinated pollen enhanced the activity. Autoradiography of the SDS-PAGE of these samples clearly showed that both actin and tubulin were substrates of TGase. Thus, the pollen TGase may be involved in the rapid cytoskeletal rearrangement which takes place during rehydration of ungerminated pollen and organization and growth of pollen tubes. Received: 9 August 1996 / Revision accepted: 26 October 1996     

Determining environmental covariates which explain genotype environment interaction in winter wheat through probe genotypes and biadditive factorial regression

Genotype-environment interaction has been analyzed in a winter-wheat breeding network using bi-additive factorial regression models. This family of models generalizes both factorial regression and biadditive (or AMMI) models; it fits especially well when abundant external information is available on genotypes and/or environments. Our approach, focused on environmental characterization, was performed with two kinds of covariates: (1) deviations of yield components measured on four probe genotypes and (2) usual indicators of yield-limiting factors. The first step was based on the analysis of a crop diagnosis on four probe genotypes. Difference of kernel number to a threshold number (DKN) and reduction of thousand-kernel weight from a potential value (RTKW) were used to characterize the grain-number formation and the grain-filling periods, respectively. Grain yield was analyzed according to a biadditive factorial regression model using eight environmental covariates (DKN and RTKW measured on each of four probe genotypes). In the second step, the usual indicators of yield-limiting factors were too numerous for the analysis of grain yield. Thus a selection of a subset of environmental covariates was performed on the analysis of DKN and RTKW for the four probe genotypes. Biadditive factorial regression models involved environmental covariates related to each deviation and included environmental main effect, sum of water deficits, an indicator of nitrogen stress, sum of daily radiation, high temperature, pressure of powdery mildew and lodging. The correlations of each environmental covariate to the synthetic variates helped to discard those poorly involved in interaction (with | correlation | <0.3). The grain yield of 12 genotypes was interpreted with the retained covariates using biadditive factorial regression. The models explained about 75% of the interaction sums of squares. In addition, the biadditive factorial regression biplot gave relevant information about the interaction of the genotypes (interaction pattern and sensitivities to environmental covariates) with respect to the environmental covariates and proved to be interesting for such an approach. Received: 8 March 1999 / Accepted: 29 July 1999     

Unexpected DNA content in the endosperm of Cupressus dupreziana A. Camus seeds and its implications in the reproductive process

 Nuclear DNA content of embryo and endosperm from mature and immature Cupressus dupreziana A. Camus seeds was estimated using laser flow cytometry. Relative DNA content of endosperm nuclei corresponded to four ploidy levels: 2C, 4C, 6C and 8C. The embryo nuclei invariably exhibited a diploid pattern. In all endosperm tissue analyzed no haploid nucleus was found. This is problematic since, in gymnosperms, endosperm and female gametes originate from one functional haploid megaspore produced by meiosis. The possible origin and derivation of C. dupreziana endosperm are discussed in light of previous results concerning the two other Mediterranean cypresses, C. sempervirens and C. atlantica. Received: 15 January 1998 / Revision accepted: 27 March 1998     

Levels of genetic diversity at different stages of the domestication cycle of interior spruce in British Columbia

Concerns over the reductionist nature of the domestication of forest-tree species focus on the possibility of potential genetic erosion during this process. To address these concerns, genetic diversity assessments in a breeding zone the Province of British Columbia “interior” spruce (Picea glauca×engelmanni) program was conducted using allozyme markers. Genetic-variation comparisons were made between natural and production (seed orchard) populations as well as seed and seedling crops produced from the same breeding zone’s seed orchard. The natural population sample consisted of a total of 360 trees representing three stands within each of three watersheds present in the Shuswap-Adams low-elevation zone of interior British Columbia. Small amounts of genetic differentiation were observed among the nine natural populations (4%) and this was attributable to extensive gene flow Consequently, the sum of these nine populations was considered as a baseline for the genetic variation present in the breeding zone. The comparisons between the seed orchard and the breeding zone produced a similar percentage of polymorphic loci while the expected hetrozygosity (0.207 vs 0.210) and the average number of alleles per locus (2.7 vs 2.4) were slightly lower in the seed orchard. A total of seven natural populations’ rare alleles were not present in the orchard population, while one allele was unique to the orchard. The %P increased to 70.6% in the seedlot, but dropped to the natural populations level (64.7%) in the plantation. The observed increase in %P was a result of pollen contamination in the orchard. It is suspected that the reduction in the plantation was caused by an unintentional selection in the nursery. Simulated roguing in the orchard did not drastically reduce even if up to 50% of the orchard’s clones were rogued. However, roguing was associated with a reduction in the average number of alleles per locus (i.e., sampling effect). Received: 2 January 1996 / Accepted: 24 May 1996     

Mapping of the Rf-3 nuclear fertility-restoring gene for WA cytoplasmic male sterility in rice using RAPD and RFLP markers

The cytoplasmic male sterility (CMS) of wild-abortive (WA) cytoplasm has been widely used for breeding hybrid rice. Two restorer genes for the CMS have been found by traditional genetic analysis. To tag the restorer genes we used a set of near-isogenic lines (NILs) of Zhenshan 97 carrying different genotypes for fertility restoration from IR24, to perform RAPD analysis. From the survey of 720 random primers, six RAPD markers were identified to be associated with Rf-3. Three of these OPK05-800, OPU10-1100 and OPW01-350, were mapped on chromosome 1. Two populations from the crosses between Zhenshan 97 A and a near-isogenic restorer line ZSR21 and between Zhenshan 97 A and IR24 were used for mapping Rf-3. The three RAPD markers and three RFLP markers, RG532, RG140 and RG458, were found to be closely linked to Rf-3 in the two populations. The same location of Rf-3 was also found in a population from the cross of IR58025 A//IR36/IR58025 B. At the RG532 locus, different alleles were found between two CMS lines, Zhenshan 97 A and IR58025 A, and between two restorer lines, IR24 and IR36. The use of these molecular markers closely linked to Rf-3 in facilitating the development of hybrid rice is discussed. Received: 3 January 1996 / Accepted: 17 May 1996     

A PCR-marker for the CMS(1) inducing cytoplasm in chives derived from recombination events affecting the mitochondrial gene atp9

The complete coding and 3′-flanking region of the mitochondrial gene atp9 of chives (Allium schoenoprasum L.) was determined in order to develop primers that allow the identification of atp9-related sequences in subsequent PCR-amplifications. One of these sequences is of a chimerical nature, consisting of atp9-homologous regions on its end, interrupted by an insertion that is composed of one atp6-homologous part and one part of unknown origin. This PCR-fragment is 762 bp in size and exclusively amplified in the sterility inducing cytoplasm of CMS₁. Thus it can be used as a PCR-marker in order to distinguish this cytoplasm type from the remaining cytoplasm types of chives. The chimerical marker sequence forms a putative open reading frame (orfA501), from which CMS₁ might originate. Received: 6 June 2001 / Accepted: 3 August 2001     

Mutations affecting pigmentation and shape of the adult zebrafish

 Mutations causing a visible phenotype in the adult serve as valuable visible genetic markers in multicellular genetic model organisms such as Drosophila melanogaster, Caenorhabditis elegans and Arabidopsis thaliana. In a large scale screen for mutations affecting early development of the zebrafish, we identified a number of mutations that are homozygous viable or semiviable. Here we describe viable mutations which produce visible phenotypes in the adult fish. These predominantly affect the fins and pigmentation, but also the eyes and body length of the adult. A number of dominant mutations caused visible phenotypes in the adult fish. Mutations in three genes, long fin, another long fin and wanda affected fin formation in the adult. Four mutations were found to cause a dominant reduction of the overall body length in the adult. The adult pigment pattern was found to be changed by dominant mutations in wanda, asterix, obelix, leopard, salz and pfeffer. Among the recessive mutations producing visible phenotypes in the homozygous adult, a group of mutations that failed to produce melanin was assayed for tyrosinase activity. Mutations in sandy produced embryos that failed to express tyrosinase activity. These are potentially useful for using tyrosinase as a marker for the generation of transgenic lines of zebrafish. Received: 17 June 1996 / Accepted: 15 July 1996     

Felinine stability in the presence of selected urine compounds

Summary. The stability of felinine, an amino acid present in feline urine, was investigated. Synthetic felinine was unstable in the urine of a selection of mammals. Felinine was found to stable in feline urine in which urea had been degraded. Synthetic felinine was found to react specifically with urea and did not react with urea analogues such as biuret or thiourea or other nucleophilic compounds such as ammonia which is more nucleophilic or acetamide and water which are less nucleophilic than urea. The reaction of urea and felinine was independent of pH over the range of 3–10. Urea did not react with N-acetyl-felinine suggesting a felinine N-terminal interaction with urea. Mass spectral analysis of the reaction products showed the presence of carbamylated felinine and fragmentation ions derived from carbamyl-felinine. The physiological relevance of felinine carbamylation is yet to be determined.     

Mapping the jp (jumbo pollen) gene and QTLs involved in multinucleate microspore formation in diploid alfalfa

The objective of this research was to map the jumbo-pollen trait in diploid alfalfa. Homozygous recessive (jpjp) plants are characterized by the complete failure of post-meiotic cytokinesis during microsporogenesis resulting in 100% 4n-pollen formation. Three F₁ segregating populations were produced and analyzed for pollen-grain production and the segregation of RFLP markers. The first cross combination did not segregate for the jumbo-pollen trait, but showed a clear segregation for multinucleate (bi-, tri- and tetra-nucleate)-microspore formation. Cytological analysis showed that few plants produced 100% normal (uninucleate) microspores, whereas most of them produced multinucleate microspores at a variable frequency (0–75%). Plants with multinucleate microspores always showed a prevalence of binucleated microspores, even though some plants showed a background presence of tri- and tetra-nucleate microspores. QTL analysis based on ANOVA I and Stepwise Multiple Regression identified three QTLs with a highly significant effect on multinucleate-microspore formation. Two cross combinations, subsequently executed, showed Mendelian segregation for the jumbo-pollen trait and were effective in locating the jp gene on linkage group 6 close to the Vg1G1b RFLP locus. Interestingly, this RFLP locus was also linked to one QTL for multinucleate-microspore formation. Genetic models are discussed concerning the presence in linkage group 6 of a cluster of genes involved in multinucleate-microspore formation together with possible relationships between the jp gene and the Vg1G1b QTL. Received: 28 January 1999 / Accepted: 16 December 1999     

Influence of soil hydric parameters on the winter cold hardiness of a burrowing beetle, Leptinotarsa decemlineata (Say)

This investigation examined the influence of soil moisture and associated parameters on the cold hardiness of the Colorado potato beetle (Leptinotarsa decemlineata Say), a temperate-zone species that overwinters in terrestrial burrows. The body mass and water content of adult beetles kept in sand at 4 °C varied over a 16-week period of diapause according to the substratum's moisture content. Changes in body water content, in turn, influenced the crystallization temperature (range −3.3 to −18.4 °C; n = 417), indicating that environmental moisture indirectly determined supercooling capacity, a measure of physiological cold hardiness. Beetles held in dry sand readily tolerated a 24-h exposure to temperatures ranging from 0° to −5 °C, but those chilled in sand containing as little as 1.7% water (dry mass) had elevated mortality. Thus, burrowing in dry soils not only promotes supercooling via its effect on water balance, but may also inhibit inoculative freezing. Mortality of beetles exposed to −5 °C for 24 h was lower in substrates composed of sand, clay and/or peat (36–52%) than in pure silica sand (78%) having an identical water content (17.0% dry mass). In addition to moisture, the texture, structure, water potential, and other physico-chemical attributes of soil may strongly influence the cold hardiness and overwintering survival of burrowing insects. Accepted: 10 September 1996     

The anterior-like cells in Dictyostelium are required for the elevation of the spores during culmination

 Shortly after initiation of Dictyostelium fruiting body formation, prespore cells begin to differentiate into non-motile spores. Although these cells lose their ability to move, they are, nevertheless, elevated to the tip of the stalk. Removal of the amoeboid anterior-like cells, located above the differentiating spores in the developing fruiting body, prevents further spore elevation although the stalk continues to elongate. Furthermore, replacement of the anterior-like cells with anterior-like cells from another fruiting body largely restores the ability to lift the spores to the top of the stalk. However, if amoeboid prestalk cells are used to replace the anterior-like cells, there is no restoration of spore elevation. Finally, when a droplet of mineral oil replaces differentiating spores, it is treated as are the spores: the mineral oil is elevated in the presence of anterior-like cells and becomes arrested on the stalk in the absence of anterior-like cells. Because a similar droplet of mineral oil is totally ignored by slug tissue, it appears that there is a dramatic transformation in the treatment of non-motile matter at this point in Dictyostelium development. Received: 26 January 1998 / Accepted: 27 May 1998     

Genetic analysis of durable leaf rust resistance in winter wheat

Quantitative resistance that delays the epidemic development of leaf rust in wheat is an important source for durable resistance breeding. The Swiss winter wheat variety ’Forno’ shows a high level of quantitative resistance against leaf rust. This resistance has been effective for more than 10 years and can therefore be considered to be durable. In order to map quantitative trait loci (QTL) for durable leaf rust resistance we analysed 204 F₅ recombinant inbred lines (RILs) of the cross between the winter wheat ’Forno’ and the winter spelt ’Oberkulmer’ for their level of leaf rust resistance (LR) and leaf tip necrosis (LTN) in four different environments. Both traits showed a continuous distribution and were significantly correlated (r=−0.5). Across environments we detected 8 QTL for leaf rust resistance (6 inherited from ’Forno’) and 10 QTL for the quantitative expression of LTN (6 inherited from ’Forno’). Of the 6 QTL responsible for the durable leaf rust resistance of ’Forno’, 1 major QTL coincided with a thaumatin locus on 7BL explaining 35% of the phenotypic variance. Four QTL for LR coincided with QTL for LTN. At these loci the alleles of ’Forno’ increased the level of resistance as well as the extent of LTN, indicating pleiotropy. Received: 1 July 1999 / Accepted: 30 July 1999     

Hibernation behavior of Rana lessonae and R. esculenta in their natural habitat

We studied the hibernation behavior of the water frog Rana lessonae and its hybridogenetic associate R. esculenta in their natural habitat during three successive winters. Animals caught in pitfall traps at a fenced pond were individually marked with PIT tags and some (n=36) were additionally equipped with radio transmitters. Of the animals caught, 85% left the fenced pond for hibernation. More R. esculenta remained inside the fenced area compared to R. lessonae. R. lessonae emigrated earlier in autumn and came back later in spring than R. esculenta, but the distance to their hibernation sites did not differ. Both species left the fenced pond earlier in the year when ambient temperatures were lower. All radio-tracked animals hibernated in woodland, 3–7 cm below the surface in soil, under moss, fallen leaves or small branches. Soil temperatures at the actual hibernation sites were significantly higher than at randomly chosen control sites. A surprising finding was that most frogs changed their hibernation sites during winter, and often more than once. Movements were more frequent in the warmer first half of the winter than in the cooler second half, but some animals were active even on days with mean temperatures below 1°C. These results show that both species do not spend the whole winter torpid in one particular hibernation site but move around, especially at higher temperatures. Most of the animals lost weight during the winter, and the weight loss was greater in females than in males and higher in warm than in cold winters. To what extent weight loss and survival is influenced by the chosen hibernation sites and the amount of movement during winter, and whether this contributes to the differences in species and sex ratios found in mixed populations, needs more investigation. Received: 4 August 1999 / Accepted: 15 November 1999